Agricultural University of Tirana, Faculty of Agriculture & Environment, Department of Horticulture and Landscape Architecture Address: Koder-Kamez, Tirana,

Albania www.ubt.edu.al

Geneva, NY, USA (March 26-28, 2014) http://events.cals.cornell.edu/ishsphysiology2014

Poster presented at the Physiological Principles and Their Application to Fruit Production

Model Unveils Changes in Stomatal Conductance in Apple Saplings after Use of Bioregulators
E. Kullaj1, V. Avdiu and F. Thomaj
1

Dep. Horticulture, Faculty of Agriculture and Environment, Agricultural University of Tirana, Kodr-Kamz, 1010, Tirana, Albania Tel: +355684096186 Email: ekullaj@ubt.edu.al

Introduction

Stomata have a major regulatory role in gas exchange in leaves and the efficiency of this regulation can impact yields of agricultural crops. Beside various environmental variables like solar irradiance, leaf temperature, leaf-to-air vapour pressure deficit (VPD), soil and plant water status, stomatal behaviour is influenced by hormones and other naturally occuring compounds. Abscisic acid (ABA) is intimately involved in the regulation of stomatal behaviour; other groups of hormones, such as the cytokinins, may also be involved in the control of stomata. Among apple cultivars, stomatal regulation has been related to tree vigour. Gibberelins/cytokinins are known to encourage shape improvement and advancing leaf apparatus emission and development. Moreover, the effects of certain environmental factors on stomatal behaviour may be mediated by hormones. However, most of reports on the effects of gibberelins and cytokinins on stomatal behaviour either refer to grasses or use excised leaves and therefore effects are not clear and not supported by most of investigators.

Material and Methods

Plant Material
Twelve standardized apple saplings bench grafted of cv. Gala Galaxy on M9 rootstocks. Plants were selected to have the same biometric features as the treated plants, namely rootstock diameter 10 cm below the grafting point and scion diameter 10 cm above the grafing point, branch number, length, crotch angle and branch diameter 5 cm from the base, number of lateral shoots, leaf area, etc. Root systems were compared as well for uniformity and size. These trees were not producing fruits. From these 12 plants planted in spring, 6 were then treated with Progerbalin LG (GA4-7 1,8% + BA 1,8%) in order to stimulate branching. The soil in which saplings were planted was of good quality, up to 60 cm deep and in average contained: humus 2.36 %, (moderate) N 0.13 % (moderate), P2O5 10.69 mg/100g soil (low), K2O 43 mg (high), Ca 101.73 mg (moderate), Mg 47.14 mg (moderate); pH value in water was 6.8 whereas in KCl 5.8 (slightly acid). Ploughing was made at 40 cm depth, organic and mineral fertilizer were distributed in advance: organic 5 kg/m2 and mineral NPK 15 : 15 : 15 100g/m2. The plot was tilled 5 times, plants were drip irrigated and have received 3 treatments with fungicides and insecticides.

The Model and the Experimental Design

Using a model(1) based on a complex parameterized Penman-Monteith equation rewritten to already include the gc as the function of R and vapour pressure deficit (VPD) (Figure 1), we have produced the dynamics of stomata conductance of these saplings, 6 treated and 2 untreated controls. (1) Transpiration data were calculated from sap flow measurements using sap flow sensors EMS 62 (EMS Brno), based on SHB
Fig. 1. (top) Script for calculating variables necessary for parametrization. (bottom) The Fitmodule in Mini32 software showing the initial parameters for the procedure of finding the model which calculates canopy transpiration and stomatal conductance of apple trees. It shows both calculated values (blue) and the actual (black) dots measured from sap flow and the correlation index (0.975)

(stem heat balance) method. Sensors were installed on shoots (12 mm thick) on 8 trees at their trunk (Figure 2). The measuring interval was every minute with 1 s warm-up and storing interval every 15 minutes during July-September 2013. A portable meteorological station Minikin RTHi (EMS Brno, CZ) measured the Rs, Ta and RH. VPD was calculated from vapour pressure and relative humidity. Plants were subject to water stress, beside others (high radiation and temperature). Soil water potential values were kept around 0,5 MPa.

The actual values of canopy transpiration (Figure 3) show a difference in average transpiration between the treated and non treated plants, with the first being constantly higher. However, daily maximum values were not statistically different. It is known for grass species only that cytokinins stimulate stomatal opening and increase rates of transpiration by delaying senescence. In turn, the delayed senescence would extend the CO2 fixing ability of the leaves resulting in lower Fig. 3. Transpiration values of treated and control apple trees leaf CO2 levels; as a consequence stomata will open more widely in the kinetin treated leaves. However, it has been suggested that the cytokinins acted on stomata indirectly by affecting water potentials elsewhere in the plant. There may also be some interaction between cytokinins, ABA and CO2 conFig. 4. Measured and calculated values of transpiration for centration. 2 2 treated (R = 0.91) and control plants (R = 0.89). Values given Almost the same goes are the average of all plants per treatment for gibberellins which have been reported to increase transpiration rates in excised leaves but most investigators find no effects in intact leaves. In epidermal strips gibberellins are also reported to have no effects on stomatal opening. Comparisons between treatments (right side) and controls (left side) of stomatal aperture and closure show changes in stomata behaviour (Figure 6). Stomata of treated trees seem to open at lower values of radiation. Even more pronounced is Fig. 5. Modeled canopy conductance of treated and control the effect on stomata clo- plants for selected days to compare changes in daily dynamics sure under high VPD. (b) Various (a) hypothesis can explain such change in stomatal behaviour. Changes in stomatal opening, although minor, could be due to the effects of bioregulators on chlorophyll content as (c) (d) the latest is the pigment mediating lightstimulated stomatal opening. Moreover, gibberellins may promote chloroplast division indirectly through impacting leaf mesophyll cell expansion, where CO2 is primarily Fig. 6. Changes in stomata behaviour, both apperture (top) and closure (bottom) between young apple trees treated with Progerbalin (right) and assimilated. Changes in stomatal closure controls (left) could be explained by the effect of gibberellins in reversing the effects of abscisic acid (ABA), as its redistribution and biosynthesis is causing stomatal closure, and its accumulation in stressed leaves plays an important role in the reduction of water loss by transpiration under water stress conditions. Further investigation will be conducted to determine the above hypothesis.

Results and Discussion

Conclusions

Fig. 2. Experimental set up and working team. Sap flow sensors on trunks of apple trees powered by solar pannels

The modeling approach using a parametrized P-M equation has unveiled a change in stomata behaviour due to the exogenous use of bioregulators, namely a combination of gibberellin A4-A7 with 6-Benzyladenine. Although the underlying mechanisms can only be hypothesized, the results represent a step forward in demonstrating that the exogenous use of bioregulators affects directly stomata behaviour.